Surface plasmon resonance imaging for real-time, label-free analysis of protein interactions with carbohydrate microarrays

Rositsa Karamanska, Jonathan Clarke, Ola Blixt, James I. MacRae, Jiquan Q. Zhang, Paul R. Crocker, Nicolas Laurent, Adam Wright, Sabine L. Flitsch, David A. Russell, Robert A. Field

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    Abstract

    Plant lectin recognition of glycans was evaluated by SPR imaging using a model array of N-biotinylated aminoethyl glycosides of beta-D-glucose ( negative control), alpha-D-mannose ( conA-responsive), beta-D-galactose ( RCA(120)-responsive) and N-acetyl-beta-D-glucosamine ( WGA-responsive) printed onto neutravidin-coated gold chips. Selective recognition of the cognate ligand was observed when RCA(120) was passed over the array surface. Limited or no binding was observed for the non-cognate ligands. SPR imaging of an array of 40 sialylated and unsialylated glycans established the binding preference of hSiglec7 for alpha 2-8-linked disialic acid structures over alpha 2-6-sialyl-LacNAcs, which in turn were recognized and bound with greater affinity than alpha 2-3-sialyl-LacNAcs. Affinity binding data could be obtained with as little as 10-20 mu g of lectin per experiment. The SPR imaging technique was also able to establish selective binding to the preferred glycan ligand when analyzing crude culture supernatant containing 10 20 mu g of recombinant hSiglec7-Fc. Our results show that SPR imaging provides results that are in agreement with those obtained from fluorescence based carbohydrate arrays but with the added advantage of label-free analysis.

    Original languageEnglish
    Pages (from-to)69-74
    Number of pages6
    JournalGlycoconjugate Journal
    Volume25
    Issue number1
    DOIs
    Publication statusPublished - Jan 2008

    Keywords

    • Carbohydrate array
    • Surface plasmon resonance (SPR) imaging
    • Label-free detection
    • Plant lectin
    • Siglec 7
    • Immunoglobulin superfamily
    • 2-bromoethyl glycosides
    • Immune system
    • Throughput
    • Binding
    • Oligosaccharides
    • Recognition
    • Lectin
    • Inhibition
    • Biosensors

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